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Objective:To explore the effects of general anesthesia and combined spinal and epidural anesthesia on inflammatory factors and pain in patients with osteoarthritis after total knee arthroplasty.Methods:A total of 84 patients with osteoarthritis who underwent unilateral total knee arthroplasty in Hulunbuir People's Hospital from January 2020 to May 2021 were selected as the research subjects. They were randomly divided into general anesthesia group (40 cases) and combined spinal and epidural anesthesia group (44 cases). Venous blood samples of 5 ml were collected before operation and 6, 24, 48 hours after operation, and the contents of inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6] in serum were determined by enzyme-linked immunosorbent assay (ELISA). The visual analogue pain scale (VAS) of the two groups at 30 min, 6, 24 and 48 hours after operation was compared.Results:At 6 and 24 hours after operation, TNF-α, IL-1β and IL-6 levels in the combined spinal and epidural anesthesia group were lower than those in the general anesthesia group ( t = 4.17, 3.85, 8.95, 10.98, 10.04, 9.87, P < 0.05). There were significant differences in the levels of TNF-α, IL-1β and IL-6 at different time points between the general anesthesia group and combined spinal and epidural anesthesia group ( F = 271.67, 149.26, 81.70, 189.36, 102.44, 157.32, P < 0.001). At 6 and 24 hours after operation, the VAS scores of patients in the combined spinal and epidural anesthesia group were significantly lower than those in the general anesthesia group ( t = 6.60, 3.66, P < 0.05). There were statistically significant differences in VSA scores between the two groups at different time points ( F = 67.47, 52.37, P < 0.05). Conclusion:The effect of combined spinal and epidural anesthesia is significantly higher than general anesthesia in inhibiting the expression of TNF-α, IL-1β and IL-6 in patients with osteoarthritis after operation, and the effect of analgesia is obvious.
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Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.
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Objective To investigate the levels of creatine kinase (CK-MB), brain natriuretic peptide (BNP), and neu-ropeptide Y (NPY) in severe pneumonia pediatric patients combined with heart failure (HF). Methods Pneumonia pediatric patients admitted from December 2010 to December 2014 had been enrolled and divided into pneumonia group (P) (n=32), se-vere pneumonia group (SP) (n=20) and severe pneumonia combined HF group (HF) (n=36). Twenty healthy children served as control group (C). Serum level of CK-MB was detected by enzyme rate method and were measured by ELISA. BNP and NPY were measured again during the recovery period in 18 cases in HF group. Results The serum levels of CK-MB, BNP and NPY were signiifcantly different among the four groups, (F=25.19~277.94, P0.05). The serum levels of BNP and NPY were not statistically signiifcant between SP and P and C group (all P>0.05). In HF group, the serum levels of CK-MB, BNP, and NPY were signiifcantly decreased in 18 cases after treatment. The serum level of BNP was positively correlat-ed with CK-MB and NPY (r=0.681, 0.525, all P<0.01) and the NPY and CK-MB levels were also positively correlated (r=0.545, P<0.01). Conclusions The Detection of the serum levels of CK-MB, BNP and NPY can help diagnose severe pneumonia com-bined with HF. The BNP maybe the most sensitive indicator.
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AIM: To investigate the molecular mechanism of Epstein-Barr virus encoded latent membrane protein 1 regulated cellular proliferation in nasopharyngeal epithelial cells. METHODS: The nasopharyngeal epithelial cells NP69 were infected with RV-pLNSX (the empty vector) and RV-LMP1 retroviruses, respectively. Therefore, the NP69-pLNSX and NP69-LMP1 cell lines were established. Sequentially, cellular proliferation of NP69-pLNSX and NP69-LMP1 cells was compared to draw the cellular growth curve. The experiments of plate clone formation and forming of soft agar colony were conducted. Meanwhile, the differential expression of proteins were identified between NP69-pLNSX and NP69-LMP1 cell lines by proteomic methods, and the expression levels of partial identified proteins were verified. RESULTS: (1) LMP1 was able to accelerate cellular proliferation of nasopharyngeal epithelial cell NP69 (n=3, P<0.05). (2) Twenty two proteins (9 up-and 13 down-regulated) of LMP1 mediated regulation were identified from infected NP69 cell lines, and the differential expression of partial identified proteins was confirmed by Western blotting and fluorescent real-time quantitative RT-PCR. CONCLUSION: LMP1 probably mediates the regulation of vimentin protein and keratin 19 protein expression to promote cellular proliferation in NP69 cells.
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To purify the extracellular domain of HER2 in vitro and improve its prokaryotic expression abundance, the cDNA fragment encoding extracellular domain of HER2 was obtained by PCR and cloned into the expression vector pGEX-6P-1. After transforming it into Escherichia coli BL21, we instituted an investigation of different inducing conditions to try out the optimal condition for expressing soluble fusion protein. As for insoluble inclusion bodies, they were dissolved in 8 M Urea and refolded in refolding buffer. The soluble protein and the refolded protein were purified with Glutathione Sepharose 4B, respectively. The results showed that both the soluble and insoluble protein existed in Escherichia coli, but the majority was insoluble. It is beneficial to the expression of soluble fusion protein by induction at lower temperature (30 degrees C) and higher optical density (A600= 1.8) with the use of certain additive in medium. By purification of the supernatant of the lysate and refolded protein, the yield of the fusion protein was about 1.23 mg per liter culture. As a result, we have obtained the maximum soluble extracellular domain of HER2 protein, and thus have laid a foundation for further work on functional study and antibody preparation for HER2.
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Humans , Cloning, Molecular , Escherichia coli , Genetics , Genes, erbB-2 , Genetics , Prokaryotic Cells , Metabolism , Protein Folding , Receptor, ErbB-2 , Genetics , Recombinant Fusion Proteins , GeneticsABSTRACT
Objective To investigate the inhibitory effect of ester catechin monomers-EGCG, GCG on the growth of human colon carcinoma cell line SW480 and its potential mechanism. Methods MTT assay, soft-agar colony formation test, Hoechst 33258 stain and flow cytometry analysis were used to determine the inhibitory effect of theasinesin(TS), and its monomers-EGCG, GCG on SW480 growth. Results EGCG, GCG and TS significantly inhibited the growth of human colon carcinoma cell line SW480 in dose-dependent manner, and their half inhibitory concentration (IC 50 ) was 108.88, 183.21, and 83.36?g?ml -1 , respectively. 24 hours after treatment with IC 50 of EGCG, GCG and TS, the colony formation rate of SW480 cells in the experimental group was obviously lower than that in the control group. Hoechst 33258 staining showed typical apoptotic features such as cell shrinkage, nuclear condensation and cell splinter in the experimental group. A subdiploid peak before G 0/G 1 phase in the experimental group was observed by flow cytometry. Conclusion In accord with TS, EGCG and GCG could inhibit the growth of colon carcinoma cell line SW480 cells, the mechanism of which may be related to inducing cell apoptosis.